Chapter title |
The Mitotic Spindle
|
---|---|
Chapter number | 22 |
Book title |
The Mitotic Spindle
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3542-0_22 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3540-6, 978-1-4939-3542-0
|
Authors |
Jones, Mathew J K, Jallepalli, Prasad V, Mathew J. K. Jones, Prasad V. Jallepalli |
Editors |
Paul Chang, Ryoma Ohi |
Abstract |
During mitosis, multiple protein kinases transform the cytoskeleton and chromosomes into new and highly dynamic structures that mediate the faithful transmission of genetic information and cell division. However, the large number and strong conservation of mammalian kinases in general pose significant obstacles to interrogating them with small molecules, due to the difficulty in identifying and validating those which are truly selective. To overcome this problem, a steric complementation strategy has been developed, in which a bulky "gatekeeper" residue within the active site of the kinase of interest is replaced with a smaller amino acid, such as glycine or alanine. The enlarged catalytic pocket can then be targeted in an allele-specific manner with bulky purine analogs. This strategy provides a general framework for dissecting kinase function with high selectivity, rapid kinetics, and reversibility. In this chapter we discuss the principles and techniques needed to implement this chemical genetic approach in mammalian cells. |
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